NOXO1 Antibody Description

Background

This antibody is designed, produced, and validated as part of a collaboration between Rockland and the National Cancer Institute (NCI) and is suitable for Cancer, Immunology and Nuclear Signaling research. The enzymes NADPH oxidase (NOX) and dual oxidase (DUOX) generate ROS in a regulated manner, producing reactive oxygen in various cells and tissues in response to growth factors, cytokines and calcium signals. The oxidase consists of the catalytic subunit gp91phox (otherwise known as NOX2), together with the regulatory subunits p22phox, p47phox, p40phox, p67phox and the small GTPase RAC. The enzyme activity of gp91phox is regulated by the assembly of these regulatory subunits with gp91phox to form an active complex. In 1999, the first of the NOX homologues of gp91phox was described as NOX1. The enzyme was cloned from a colon epithelial cell complementary DNA library. When expressed in cells, NOX1 generated low amounts of ROS, but high-level ROS production by NOX1 was subsequently achieved by co-expression with novel regulatory subunits (described later). Subsequently, NOX3 and NOX4 were cloned, and the latter was shown to generate high levels of ROS when expressed in cells. NOX organizer 1 (NOXO1) is a homologue of p47phox and has an almost identical domain organization, except that it lacks the auto-inhibitory region. NOX activator 1 (NOXA1) is a homologue of p67phox and similarly shares the same domain organization, except that it lacks one of the two SH3 domains that are present in p67phox. Co-transfection of NOX1, NOXO1 and NOXA1 results in marked ROS generation. Similar to p47phox, NOXO1 binds to p22phox, which is required for NOX1-dependent activity. NOXA1 has a well-conserved activation domain, implying a conserved mechanism for regulating the activity of the target NOX enzyme.

Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.

Application Note

Anti-NOXO1 antibody has been tested for use in ELISA, immunohistochemistry and western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 50 kDa in size corresponding to NOXO1 protein by western blotting in the appropriate cell lysate or extract.

Purity/Specifity

Anti-NOXO-1 antibody is directed against human NOXO1 protein. The product was affinity purified from monospecific antiserum by immunoaffinity purification. A BLAST analysis was used to suggest cross reactivity with NOXO1 protein from human and chimpanzee (100% homology). Expect reactivity with alpha, delta and gamma isoforms of NOXO1. Also expect partial reactivity against NOXO1 homologues from dog and guinea pig (87%), as well as rat (75%) and mouse (68%). Reactivity against homologues from other sources is not known.

Disclaimer Note-General

This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.